Effect Of GNB2L1 On Proliferation And Migration Of Malignant Melanoma Cells

Background:Malignant melanoma(MM)is a highly malignant tumor produced by the abnormal proliferation of melanocytes.In recent years,the high metastatic nature of malignant melanoma has been a difficult point in its treatment.At present,many studies have reported that GNB2L1 acts as a multifunctional scaffold protein,mediates the transmission of multiple signaling pathways in cells,and participates in various tumorigenesis,proliferation,invasion and migration processes.However,the effects of the GNB2L1 on various biological behaviors of malignant melanoma have not been reported,especially for the proliferation and migration of malignant melanoma.In this study,GNB2L1 specific sequence RNA interference lentivirus was used to silence the expression of GNB2L1 protein,and then the effect of GNB2L1 protein silencing on proliferation and migration of mouse melanoma B16F10 cells was observed.Objective:After inhibiting the expression of GNB2L1 by silencing GNB2L1,the proliferation and migration of mouse melanoma B16F10 cells were observed.The effect of GNB2L1 expression level on the biological behavior of mouse malignant melanoma cell line B16F10 was studied in depth,which was the clinical treatment of GNB2L1 as malignant melanoma.,Provide theoretical basis and guiding significance for the clinical treatment of GNB2L1 as malignant melanoma.Methods:1.Western blot was used to detect the expression of GNB2L1 in human melanoma A375,A2058 cells and mouse melanoma B16F10 cells.2.Using mouse melanoma B16F10 cells as the research object,The specific sequence and irrelevant sequence targeting the GNB2L1 locus were designed to make GNB2L1 interfering lentiviral solution and empty vector lentiviral solution,and stably infected with Mouse melanoma B16F10 cells to obtain respectively B16F10-GNB2L1-shRNA cells targeting GNB2L1 locus(experimental group)and unrelated sequence B16F10-NC cells(negative control group),uninfected B16F10 cells were set as blank control group.The expression of GNB2L1 protein in B16F10-GNB2L1-shRNA cells and B16F10-NC cells was detected by Western Blot at the protein level,and uninfected B16F10 cells were used as blank controls.3.Western Blot was used to detect the expression levels of epithelial-mesenchymal transition(EMT)proteins marker E-cadherin and N-cadherin in cells B16F10-GNB2L1-shRNA and B16F10-NC,and the uninfected cells B16F10 were used as blank controls.4.The CCK-8 method was used to detect respectively the proliferation of B16F10-GNB2L1-shRNA and B16F10-NC,and the uninfected cells B16F10 were used as blank controls.5.The scratch test was used to detect respectively the migration ability of B16F10-GNB2L1-shRNA and B16F10-NC,and the uninfected cells B16F10 were used as blank controls.Results:1.Western Blot results showed that GNB2L1 protein was highly expressed in human melanoma cells A375,A2058 and murine melanoma cells B16F10.2.Western Blot showed that compared with the blank group B16F10 and the negative control group B16F10-NC,the expression of GNB2L1 protein in B16F10-GNB2L1-shRNA was significantly decreased(P<0.05);The expression of GNB2L1 protein in B16F10-NC cells and B16F10 cells have no obvious difference(P>0.05).3.Western Blot showed that compared with the blank group B16F10 and the negative control group B16F10-NC,the expression of E-cadherin protein in B16F10 stably transfected cell B16F10-GNB2L1-shRNA was significantly increased(P <0.05),The expression of E-cadherin protein in B16F10-NC cells and B16F10 cells have no obvious difference(P > 0.05);Compared with blank group B16F10 and negative control group B16F10-NC,the expression of N-cadherin protein in B16F10 stably transfected cell B16F10-GNB2L1-shRNA was significantly decreased(P <0.05).The expression of N-cadherin protein in B16F10-NC cells and B16F10 cells have no obvious difference(P>0.05).4.The results of CCK-8 showed that compared with the blank group B16F10 and the negative control group B16F10-NC,the proliferation activity of B16F10-GNB2L1-shRNA was significantly decreased(P<0.05);The proliferation activity of B16F10-NC cells and B16F10 cells have no obvious difference(P>0.05).5.Scratch test proved that compared with the blank group B16F10 and the negative control group B16F10-NC,the migration ability of B16F10-GNB2L1-shRNA was significantly reduced(P<0.05);The migration ability of B16F10-NC cells and B16F10 cells have no obvious difference(P>0.05).Conclusions:1.Lentiviral vector-mediated short hairpin RNA(shRNA)can effectively silence the expression of GNB2L1 protein in mouse melanoma B16F10 cells.2.Construction of B16F10 stably transfected B16F10-GNB2L1-shRNA cells can effectively increase the expression of E-cadherin protein and decrease the expression of N-cadherin protein.3.Down-regulation of GNB2L1 protein expression can effectively slow the proliferation of mouse melanoma B16F10 cells and reduce the migration ability of cells.It is confirmed that GNB2L1 plays an important role in regulating the proliferation,migration and epithelialization of malignant melanoma.GNB2L1 is expected to be an effective target for the treatment of malignant melanoma.